biostudies-literatureUnknownZhou XSwiss National Science FoundationNIDA NIH HHSNIMH NIH HHSNociceptin/orphanin-FQ (N/OFQ) is a recently appreciated critical opioid peptide with key regulatory functions in several central behavioral processes including motivation, stress, feeding, and sleep. The functional relevance of N/OFQ action in the mammalian brain remains unclear due to a lack of high-resolution approaches to detect this neuropeptide with appropriate spatial and temporal resolution. Here we develop and characterize NOPLight, a genetically encoded sensor that sensitively reports changes in endogenous N/OFQ release. We characterized the affinity, pharmacological profile, spectral properties, kinetics, ligand selectivity, and potential interaction with intracellular signal transducers of NOPLight <i>in vitro</i>. Its functionality was established in acute brain slices by exogeneous N/OFQ application and chemogenetic induction of endogenous N/OFQ release from PNOC neurons. <i>In vivo</i> studies with fibre photometry enabled direct recording of NOPLight binding to exogenous N/OFQ receptor ligands, as well as detection of endogenous N/OFQ release within the paranigral ventral tegmental area (pnVTA) during natural behaviors and chemogenetic activation of PNOC neurons. In summary, we show here that NOPLight can be used to detect N/OFQ opioid peptide signal dynamics in tissue and freely behaving animals.bioRxiv : the preprint server for biology2023.05.26.542102https://www.ebi.ac.uk/biostudies/studies/S-EPMC10245933biostudies-literatureDevelopment of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release.PMC10245933P50 MH119467310030F31 DA059438196455T32 DA007278Suko ABhat MAAchanta ASPalmiter RDJohnson JCBruning JCDernic JRavotto LZhou XAssoumou KWeber BStine CSteuernagel LBauder CAPasqualini ALBruchas MRFusca DKloppenburg PPrada POJadhav SBenke DPatriarchi TStoeber MfalseDevelopment of a genetically encoded sensor for probing endogenous nociceptin opioid peptide release.Nociceptin/orphanin-FQ (N/OFQ) is a recently appreciated critical opioid peptide with key regulatory functions in several central behavioral processes including motivation, stress, feeding, and sleep. The functional relevance of N/OFQ action in the mammalian brain remains unclear due to a lack of high-resolution approaches to detect this neuropeptide with appropriate spatial and temporal resolution. Here we develop and characterize NOPLight, a genetically encoded sensor that sensitively reports changes in endogenous N/OFQ release. We characterized the affinity, pharmacological profile, spectral properties, kinetics, ligand selectivity, and potential interaction with intracellular signal transducers of NOPLight <i>in vitro</i>. Its functionality was established in acute brain slices by exogeneous N/OFQ application and chemogenetic induction of endogenous N/OFQ release from PNOC neurons. <i>In vivo</i> studies with fibre photometry enabled direct recording of NOPLight binding to exogenous N/OFQ receptor ligands, as well as detection of endogenous N/OFQ release within the paranigral ventral tegmental area (pnVTA) during natural behaviors and chemogenetic activation of PNOC neurons. In summary, we show here that NOPLight can be used to detect N/OFQ opioid peptide signal dynamics in tissue and freely behaving animals.2024-01-01T00:00:00Z2024 May2026-04-08T14:11:50.831Z2025-07-12T03:06:10.877ZS-EPMC102459333729295710.1101/2023.05.26.542102